Kavi Harsh H, Birchler James A
Division of Biological Sciences, University of Missouri, Columbia, MO-65211, USA.
BMC Res Notes. 2009 Oct 23;2:217. doi: 10.1186/1756-0500-2-217.
CG11033 (dKDM2) is the Drosophila homolog of the gene KDM2B. dKDM2 has been known to possess histone lysine demethylase activity towards H3K36me2 in cell lines and it regulates H2A ubiquitination. The human homolog of the gene has dual activity towards H3K36me2 as well as H3K4me3, and plays an important role in cellular senescence.
We have used transgenic flies bearing an RNAi construct for the dKDM2 gene. The knockdown of dKDM2 gene was performed by crossing UAS-RNAi-dKDM2 flies with actin-Gal4 flies. Western blots of acid extracted histones and immunofluoresence analysis of polytene chromosome showed the activity of the enzyme dKDM2 to be specific for H3K4me3 in adult flies. Immunofluoresence analysis of polytene chromosome also revealed the presence of multiple nucleoli in RNAi knockdown mutants of dKDM2 and decreased H3-acetylation marks associated with active transcription.
Our findings indicate that dKDM2 is a histone lysine demethylase with specificity for H3K4me3 and regulates nucleolar organization.
CG11033(dKDM2)是基因KDM2B的果蝇同源物。已知dKDM2在细胞系中对H3K36me2具有组蛋白赖氨酸去甲基化酶活性,并且它调节H2A泛素化。该基因的人类同源物对H3K36me2以及H3K4me3具有双重活性,并在细胞衰老中起重要作用。
我们使用了携带dKDM2基因RNAi构建体的转基因果蝇。通过将UAS-RNAi-dKDM2果蝇与肌动蛋白-Gal4果蝇杂交来实现dKDM2基因的敲低。酸性提取组蛋白的蛋白质免疫印迹和多线染色体的免疫荧光分析表明,dKDM2酶的活性在成年果蝇中对H3K4me3具有特异性。多线染色体的免疫荧光分析还揭示了dKDM2的RNAi敲低突变体中存在多个核仁,以及与活跃转录相关的H3-乙酰化标记减少。
我们的研究结果表明,dKDM2是一种对H3K4me3具有特异性的组蛋白赖氨酸去甲基化酶,并调节核仁组织。